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尖孢镰刀菌在与寄主的相互作用中分泌几个特定的富含半胱氨酸的小分子量蛋白进入木质部中启动致病力,被称为SIX(secreted in xylem)蛋白,为明确其在不同寄主中的作用,本研究比较分析了几种尖孢镰刀菌专化型中SIX1、SIX4、SIX6、SIX8同源基因序列。根据已完成的尖孢镰刀菌古巴专化型1号(Foc1)与4号生理小种(Foc4)全基因组测序序列信息及相关SIX基因序列设计引物,应用PCR方法扩增分析56株尖孢镰刀菌古巴专化型与18株其它专化型及非致病型尖孢镰刀菌菌与其它种或属共21株菌株中的SIX1、SIX4、SIX6、SIX8基因。结果表明:设计的SIX1、SIX4、SIX6、SIX8基因引物均不能从非致病性尖孢镰刀菌与其它镰刀属种或其它属的菌株DNA中扩增出目的条带;SIX1基因的2个引物均能从供试的Foc菌株DNA中扩增出目的条带,同时可从部分其它专化型菌株DNA中扩增出目的条带;SIX4基因引物仅能从供试的尖孢镰刀菌番茄专化型与部分甘蓝专化型菌株DNA中扩增出目的条带;SIX6引物仅能从供试Foc1、Foc2、Foc4菌株DNA中扩增出目的条带;SIX8基因引物能从所有供试的致病尖孢镰刀菌中DNA扩增出目的条带。研究发现的SIX6基因序列提供了快速鉴定尖孢镰刀菌古巴专化型的检测方法,同时为深入研究尖孢镰刀菌各个专化型中SIX基因的功能奠定基础。
Fusarium oxysporum secretes several specific small cysteine-rich proteins into the xylem to initiate virulence in the interaction with the host, called SIX (secreted in xylem) protein, Host, the present study compared and analyzed the sequences of SIX1, SIX4, SIX6 and SIX8 homologous genes in Fusarium oxysporum f. Primers were designed based on the complete sequence information of Foc1 and Foc4 genome sequence of Foci and the related SIX gene sequences. PCR was used to amplify and analyze 56 strains of Fusarium oxysporum SIX1, SIX4, SIX6 and SIX8 genes in 21 strains of Fusarium of Cuba, 18 strains of Fusarium isolates of non-pathogenic Fusarium oxysporum and other species or genus. The results showed that the primers of SIX1, SIX4, SIX6 and SIX8 were not able to amplify the target DNA from non-pathogenic Fusarium oxysporum and other species of Fusarium or other genus. The two primers of SIX1 gene The target bands could be amplified from the DNA of the tested Foc strains and the target bands could be amplified from DNA of some other special strains. The SIX4 gene primers could only be amplified from Fusarium oxysporum PCR-RFLP, PCR-RFLP and PCR-RFLP. The target bands were amplified from the DNA of the genotypes and some of the cabbage strains. The SIX6 primers only amplified the target DNA from the Foc1, Foc2 and Foc4 strains. Fusarium oxysporum DNA amplification of the purpose of the band. The SIX6 gene sequence found in the study provided a rapid method for the detection of Fusarium oxysporum fusiformis and laid the foundation for further study on the function of SIX gene in various specialized Fusarium oxysporum species.